| 英文摘要 | 第8页 |
| 中文摘要 | 第10-12页 |
| Chapter 1 REVIEW | 第12-28页 |
| 1．1 Biodegradation Of Crude Oil Hydrocarbon By Microorganism | 第12-15页 |
| 1．1．1 Chemical and physical characteristics of crude oil hydrocarbons | 第12-13页 |
| 1．1．2 Hydrocarbon degrading microorganisms | 第13-14页 |
| 1．1．3 Bioremediation of crude oil contamination | 第14-15页 |
| 1．2 Phylogenetic Analysis of Bacteria Based on 16S rDNA and ITS | 第15-20页 |
| 1．2．1 16S rDNA of bacteria | 第16-19页 |
| 1．2．1．1 PCR amplification of 16S rDNA | 第17-18页 |
| 1．2．1．2 Generation of phylogenetic tree using 16S rDNA sequences | 第18-19页 |
| 1．2．2 ITS of bacteria | 第19-20页 |
| 1．3 Bioinformatic resources for phylogenetic analysis of Bacteria | 第20-22页 |
| 1．3．1 Free software for phylogenetic analysis | 第20-21页 |
| 1．3．1．1 ClustalX | 第20-21页 |
| 1．3．1．2 PHYLIP | 第21页 |
| 1．3．2 Phylogenetic analysis online | 第21-22页 |
| 1．3．2．1 BLAST | 第21页 |
| 1．3．2．2 RDP | 第21页 |
| 1．3．2．3 SSU rRNA | 第21-22页 |
| 1．4 Application of 16S rDNA/ITS-Based Methods in Microbial Ecology | 第22-28页 |
| 1．4．1 Microbial diversity of hydrocarbon degrading bacteda | 第22-23页 |
| 1．4．2 Specific detection of hydrocarbon degrading bacteria using 16S rDNA | 第23-25页 |
| 1．4．3 Amplified ribosomal DNA restriction analysis （ARDRA） | 第25页 |
| 1．4．4 ITS length polymorphism | 第25-26页 |
| 1．4．5 Application of DNA microarray to microbial ecology | 第26-28页 |
| Chapter 2 Isolation and Characteristics of Crude Oil-Degrading Bacteria | 第28-42页 |
| 2．1 Materials and Methods | 第28-32页 |
| 2．1．1 Chemicals | 第28页 |
| 2．1．2 Sample sources | 第28页 |
| 2．1．3 Media for isolation of hydrocarbon degrading bacteria | 第28-29页 |
| 2．1．4 Procedures for isolation of hydrocarbon-degrading bacteria | 第29页 |
| 2．1．5 Microscopic examinatio | 第29页 |
| 2．1．6 Characteristics of isolates by BioMerieux Vitek Microbiological Identification Syste | 第29-32页 |
| 2．1．6．1 BioMerieux Vitek Microbiological Identification System | 第30页 |
| 2．1．6．2 Procedure of identification by using GNI card | 第30-32页 |
| 2．1．7 Utility of carbon sources | 第32页 |
| 2．1．8 Biodegrading characteristics of isolates | 第32页 |
| 2．2 Results and Analysis | 第32-42页 |
| 2．2．1 Isolation of crude oil degrading bacteria | 第32-33页 |
| 2．2．2 Morphological characteristics of hydrocarbon degrading bacteria | 第33-34页 |
| 2．2．3 Biochemical Characteristics of hydrocarbon-degrading bacteda | 第34-36页 |
| 2．2．4 Utilization of Hydrocarbon | 第36-40页 |
| 2．2．5 Degrading characteristics of hydrocarbon by isolates | 第40-41页 |
| 2．2．6 Isolation and Morphology of crude oil-degrading fungus | 第41-42页 |
| Chapter 3 Phylogenetic Analysis of Crude Oil-Degrading Bacteria | 第42-51页 |
| 3．1 Materials and Methods | 第42-43页 |
| 3．1．1 Primers | 第42页 |
| 3．1．2 PCR-Amplification of 16S rDNA | 第42-43页 |
| 3．1．3 Sequences alignment and phylogenetic analysis | 第43页 |
| 3．2 Results and Analysis | 第43-51页 |
| 3．2．1 Sequences alignment of 16S rDNA | 第43-44页 |
| 3．2．2 Phylogenetic analysis based on partial sequence of 16S rDNA | 第44-47页 |
| 3．2．3 Phylogenetic analysis of Acinetobacter isolates | 第47-51页 |
| Chapter 4 ITS Analysis of Crude oil-Degrading Bacteria | 第51-68页 |
| 4．1 Materials and Methods | 第51-52页 |
| 4．1．1 PCR-Amplification of ITS | 第51-52页 |
| 4．1．2 Sequences alignment and phylogenetic analysis | 第52页 |
| 4．2 Results and Analysis | 第52-68页 |
| 4．2．1 PCR-Amplification of ITS using different pairs of primer | 第53-55页 |
| 4．2．2 Sequence alignment and comparative analysis of ITS | 第55页 |
| 4．2．3 tRNA sequence identified in the 16S-23S ITS region | 第55-65页 |
| 4．2．4 Phylogenetic tree based on ITS of hydrocarbon-degrading isolates | 第65-68页 |
| Chapter 5 Conclusion and Perspective | 第68-74页 |
| 5．1 Isolation and characteristics of hydrocarbon-degrading bacteria | 第68页 |
| 5．2 PCR-amplificafion of 16S rDNA and ITS | 第68-69页 |
| 5．2．1 Precautionary measures | 第69页 |
| 5．2．2 Preparation of bacterial DNA Template | 第69页 |
| 5．3 Pseudomonas aeruginosa: degradative strains and pathogenic strains | 第69-71页 |
| 5．4 Classification of isolates belonging to Acinetobacter | 第71-73页 |
| 5．4．1 Classification of strains in Acinetobacter | 第71-72页 |
| 5．4．2 Differentiation of Acinetobacter strains by molecular methods | 第72-73页 |
| 5．5 Identification of bacteria: from morphology to 16S rDNA-ITS sequence | 第73-74页 |
| References | 第74-80页 |
| Acknowledgements | 第80-81页 |
